7th Asia-Pacific Conference on Plasma Physics, 12-17 Nov, 2023 at Port Messe Nagoya

Non-Thermal Atmospheric Pressure Plasma for Controlling Cell Fate
Shinya Kumagai1
1Department of Electrical and Electronic Engineering, Meijo University
e-mail: skumagai@meijo-u.ac.jp

Non-thermal atmospheric pressure plasma (NTAPP) has
been used in biomedical research [1-5]. One of theattractive applications is gene transfection for controllingcell fate [6,7]. Compared to conventional genetransfection methods using virus vector, lipofection, orelectroporation, non-thermal atmospheric pressureplasma is expected to achieve superior gene transfection
without causing damages to cells. Excellent results havebeen reported but what is happening during plasma genetransfection at each cell has been unclear. In this study,
surface morphology of cells upon plasma exposure wereanalyzed in detail. The plasma-induced surface structure
was discussed in terms of gene transfer into cells.
In the plasma gene transfection, cells cultured in a dishis exposed to plasma. Plasma exposure can increase thematerial permeability of the cell membrane enabling
gene transfer into cells (Fig. 1).
To elucidate plasma exposure effects on cell membrane,cells exposed to plasma jet were freeze-dried andanalyzed with Scanning Electron Microscope (SEM).
SEM observation revealed that cell membrane before
plasma exposure (control sample) had smooth but wavystructures. At the beginning of plasma exposure,conditions of cell membrane were almost the same. With
increasing the plasma exposure time, protruded
structures formed on the cell membrane. With further
increasing, it was found that nanopores formed on thecell membrane.
Towards the gene transfer, substance intake experiment
was conducted. Fluorescent reagent of DiYO-1,(molecular weight: 1270) was used. Compared to using
plasmid DNA for introduction inside cells, merit of using
fluorescent reagent is that introduction of fluorescentreagent can be analyzed easily by fluorescencemicroscopy without incubating cells for gene expression.
For the cell samples in the beginning of plasma
exposure, fluorescence of DiYO-1 was not observed.
With increasing plasma exposure time, fluorescent wasobserved for the cells under plasma exposure conditionsof nanopore formation.
Considering these results obtained here, nanopores

formed by plasma can play important roles in substanceintake.
GlasstubeElectrodePlasma jetActivespecies

He gasHigh voltage
sourceDNAPlasmaeffects

2. Gene transfection throughpermeability-improved cellmembrane
1. Plasma exposure

Fig. 1 Plasma gene transfection.

Acknowledgement
This study was partially supported by JSPS KAKENHI(Scientific Research (B) 19H04457; Challenging
Exploratory Research 18K19942, 26600130); The
Takahashi Industrial and Economic ResearchFoundation; The joint usage/research program of centerfor Low-temperature Plasma Sciences, Nagoya
University.
References[1] Ishikawa et al., Free Radical Research 57, 239(2023),
https://doi.org/10.1080/10715762.2023.2230351[2] H. Tanaka et al., Sci. Rep. 9, 13657 (2019).
[3] Y. Hori et al., Plasma Process Polym. 18, e2000225(2021).
[4] M. Kobayashi et al., Heliyon 2022,
https://doi.org/10.1016/j.heliyon.2022.e12009(OPEN ACCESS).
[5] S. Kumagai et al., JSAP Rev. 2022, 220417,
https://doi.org/10.11470/jsaprev.220417(OPEN ACCESS).
[6] Ogawa et al., Biotechnol. Bioeng. 92, 865 (2005).
[7] Jinno et al., Plasma Sources Sci. Technol. 26, 6,(2017).